ExiProgen™ EC1 Protein Synthesis Kit

ExiProgen™ EC1 Protein Synthesis Kit is designed for synthesis and
purification of high-quality protein in under 6 hours —
simply add template DNA.

Bioneer's ExiProgen™ EC1 Protein Synthesis Kit is designed for in vitro (cell-free) expression and high-yield Histidine-Tag affinity purification of proteins by using the world's first automated protein synthesis and nucleic acid extraction system "ExiProgen™". 1 to 16 different types of highly pure proteins can be obtained from each run. Each reaction needs only 6-10 µg of DNA, which can be in the form of an expression vector such as Bioneer's pBIVT or a PCR product using the ExiProgen™ ProXpress PCR Template Kit. Because the entire process is automated, you will obtain reliable and reproducible results run after run.

The template DNA for expression by ExiProgen™ EC1 Protein Synthesis Kit must have a "T7 promoter – ribosomal biding site (RBS) – target gene – T7 terminator" structure, and target gene must contain a histidine tag coding sequence at N-terminus or C-terminus for the protein purification.

Maximization of Protein Expression

Protein can be normally synthesized with 6-10 µg of template DNA, but the yield of protein synthesized in the reaction depends on the amount of DNA used for expression. Therefore, a preliminary experiment described as below is recommended to determine the amount of template DNA to maximize the expression of each protein using the AccuRapid™ Cell-free Protein Expression Kit. Note that more DNA is not always better!

Example: Synthesis of poly(A) polymerase

1. Determination of the optimal amount of template DNA for the expression of poly(A) polymerase using AccuRapid™ Cell-Free Protein Expression Kit
; Poly(A) polymerase was maximally expressed with the AccuRapid™ kit when 400 ng of pET-poly(A) polymerase was used.

1. Reproducibility:ExiProgen™ EC1 Protein Synthesis Kits provide reproducible results, the first time and every time.Figure 1. Expression and purification of CAT. Reproducibility is seen in all 16 wells with no detectable variation between wells.
Lane 1~16: Number of the well,
M : AccuLadder™ Protein Size Marker (Low),
E : Expression sample,
P : Purification sample

2. Flexibility: Proteins as small as 10 kD and as large as 120 kD have been expressed/purified with ExiProgen™.

Figure 2. Expression and purification of various proteins. Up to 16 types of proteins can be expressed and purified simultaneously with an average of over 90% purity.
Top of the lane: Name of the protein,
M : AccuLadder™ Protein Size Marker (Low),
E : Expression sample,
P : Purification sample

3. Control: Each kit comes with a Green Fluorescent Protein positive control providing confidence in the results.Figure 3. Detection of fluorescence emitted from fluorescent proteins synthesized with ExiProgen™.
This result indicates that these synthesized SfGFP, RFP, and AcGFP are functionally active.
Left panel: Color of each protein elution samples detected with naked eyes.
Right panel: Fluorescence from protein elution samples detected with UV illuminator.

4. Functionality: Proteins expressed/purified with ExiProgen™ can be assayed for functionality and the resulting proteins will generally yield up to 100 µg or > 90% pure protein. Many established recombinant proteins can be expressed in E. coli extracts with full functional activity.